Use of dextran and post-primary antibody fixation in immunoperoxidase staining of fresh frozen tissue. Detection of immunoglobulin associated with squamous carcinomas of the head and neck.

Abstract

Use of unfixed fresh frozen tissue sections for immunocytochemical studies reduces the possibility of denaturation of antigenic determinants compared to formalin fixation and paraffin embedding procedures. However, tissue and cellular morphology can be extensively altered in the numerous application and washing steps with frozen tissue sections. We tested a number of buffer solutions and showed that the use of dextran-containing buffers and fixation by glutaraldehyde after primary antibody application preserves tissue morphology. The procedures described here are also applicable to ascertaining the presence of Fc receptors of leukocytes in sections of carcinoma tissues. The buffered dextran washes and post-primary antibody fixation method was used to demonstrate the presence of immunoglobulin associated with squamous carcinoma cells. The immunoglobulin was not removed by washing of tissue sections at 37 degrees C but could be removed by low or high pH buffer washes, suggesting that the immunoglobulin is bound in a specific manner.